LOCUS       sequence-413327-        4050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Riboprobe generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4050)
  AUTHORS   Park SJ, Silic MR, Staab PL, Chen J, Zackschewski EL, Zhang G
  TITLE     Evolution of two-pore domain potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2024 Jan 25. doi: 10.1002/dvdy.690.
  PUBMED    38270285
REFERENCE   2  (bases 1 to 4050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Dyn. 
            2024 Jan 25. doi: 10.1002/dvdy.690."
COMMENT     Sequence Label: pENTR-D-Topo-kcnk9
FEATURES             Location/Qualifiers
     source          1..4050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..28
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     20..37
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     21..37
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    53..152
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1659..1758)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1775..1794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1776..1794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1799..1815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1799..1815)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1917..1936)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1928..2737
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1995..2014)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2884..3472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3373..3392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3626..3643
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3802..3829)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3921..4007)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
       61 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      121 tataatgcca actttgtaca aaaaagcagg ctccgcggcc gcccccttca ccatggcact
      181 gcgcgccggg aggactttgc tgtggttcgg gcaggttttg cgcagagttt cgcggctgca
      241 gggcttgtgg tcccggcgct ccagcagtct gctgtgtctc tccaacagcc aggagcacga
      301 ccaagggacc tgttacagcc gcaaccggtg tccgaagacg catcggcagc aacactgccc
      361 ctatccgccc tttccagact gctgctgcgc cttcccgggg gataagaggg gacatgaacc
      421 cctcagccgc cggttcctgg tggccatgaa gaggcagaac gtgcggacgc tgtccctgat
      481 cgtctgcacg ttcacctatc tgctggtcgg ggccgcggtg tttgacgcgc tcgagtcgga
      541 cttcgagatg cgggagaagg agcagctgga ggcggaggag aagcgcctcc agggcaagta
      601 caacatcagt gaggatgact ataaaaagct gcagaccatc atcatggagg ccgagccgca
      661 cagagcaggg gtccagtgga aatttgcagg ctcgttttac tttgccatca ctgtcataac
      721 cactataggt tatggtcacg cagctccagg cacagatgct gggaaggcct tctgcatgtt
      781 ttacgccgtg ctggggatcc cgctcacgct ggtgatgttc cagagcctcg gcgagcggat
      841 gaacaccttc gtcaagtact tgctaaagcg catcaagaag tgctgcggga tgcgcatcac
      901 agaggtgtcc atggagaaca tggtaacagt gggcttcttc tcttgcatgg gcacgctctg
      961 catcggcgct gccgcatttt cccagtacga ggactggagc ttcttccagt cctattacta
     1021 ttgtttcata acactcacca ctattggctt cggggacttc gtggccctcc agaagaacaa
     1081 agccctgcag aaaaagcctc tttacgtggc gttcagcttc atgtacatcc tagtggggct
     1141 gacggtcatc ggggccttcc tcaatctggt ggtgctacgc ttcttgacca tgaacagcga
     1201 ggacgagcga cgggatgctg aggagagggc gtccctggcg gggaaccgca gtagcatgat
     1261 catccacatc caagaagaca cgcttcagag gagtcggcgg aggcgagaac agcagcagaa
     1321 ccgctaccgt cccgaagtca ccgatctcca gtcggtttgc tcctgcatgc actaccactc
     1381 gcatgagttc gggagctcgg tcggcctcgg gttgggattg ggaggaggtg gcggaggtgc
     1441 gtttccgcag cagaactcat tcggctcgca gttgagcccg catccgtacc atcactacca
     1501 ctcaacggtg tcctaccgca tcgaggagat ctcaccgagc accttgaaaa acagcttcct
     1561 tccttccccg atcagctcca tctctcctgg gctgcacagc ttcgcggaaa acctccggct
     1621 tatgagacga cgcaagtcca tcaagggtgg gcgcgccgac ccagctttct tgtacaaagt
     1681 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1741 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1801 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1861 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1921 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1981 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2041 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2101 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2161 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2221 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2281 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2341 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2401 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2461 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2521 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2581 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2641 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2701 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2761 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2821 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2881 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2941 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3001 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3061 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3121 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3181 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3241 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3301 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3361 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3421 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3481 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3541 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3601 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3661 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3721 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     3781 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     3841 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     3901 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     3961 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     4021 ccctactctc gcgttaacgc tagcatggat
//
